Do Psychostimulatory Drugs Enhance Lentivirus Infection?

描述(由申请人提供)：在过去十年中，甲基苯丙胺(冰毒)的使用与艾滋病毒-1感染的发病率上升有关。此外，已报告的在冰毒使用者中迅速发展的艾滋病毒-1感染病例强调了确定冰毒对艾滋病毒-1感染中枢神经系统的影响的重要性。这笔赠款的目的是检验这一假设，即冰毒(以及可能的其他精神刺激药物)通过加速感染事件的频率和伴随的反转录错误的发生率来增强艾滋病毒抗病毒药物耐药突变的发展。艾滋病毒耐药基因型的出现容易发生在中枢神经系统(CNS)，那里的药物浓度可能不是有效控制艾滋病毒复制的最佳药物浓度。与此同时，由于血脑屏障(BBB)对T细胞贩运的限制，中枢神经系统被认为受到免疫系统的较少监控。因此，中枢神经系统是艾滋病毒感染、复制、适应和选择的特权场所。此外，中枢神经系统可能是耐药变异体进入外周隔室的储存库。该提案有两个具体目标。具体目标1是验证冰毒在体外加速耐药慢病毒发展的假设。这个目标有四个目标：#1：比较在存在或不存在冰毒的情况下培养的星形胶质细胞和小胶质细胞对AZT耐药的FIV的发展速度。目的#2：比较有或无冰毒培养的星形胶质细胞和小胶质细胞对AZT耐药HIV的体外发育率。目的#3：检测冰毒对未感染和慢病毒感染的淋巴细胞、星形胶质细胞和小胶质细胞产生炎性细胞因子的影响及其与病毒表达的关系。目的#4：确定冰毒是否增强HIV对人类靶细胞的感染，促进细胞增殖，并调节CXCR4和/或CCR5的表达。具体目的#2旨在通过在FIV/CAT系统中进行原理验证研究，在体内验证这一假说，以评估冰毒对体内AZT耐药性的影响。精神刺激药物，包括广泛的合法和非法物质，可以改变艾滋病毒的发病机制并影响耐药变种的出现，这一前景是新的。以前有人提出，隔离在中枢神经系统的艾滋病毒可能有更大的抗药性余地，但冰毒在这一过程中可能发挥的作用是这一概念的一个新方面。项目简介：中枢神经系统是慢病毒感染的蓄水池，有别于外周隔室，允许病毒变体，特别是抗病毒耐药突变株的独立进化。根据我们的观察，我们推测冰毒可能会促进中枢神经系统中耐药病毒的发展，然后这些病毒可以进入外周间隔。最近的一份报告显示，纽约一名冰毒使用者感染了一种快速发展的、高度耐药的艾滋病毒-1病毒，这突显了阐明冰毒与艾滋病毒-1之间的相互作用以及中枢神经系统耐药发展的重要性。此外，冰毒是可能对中枢神经系统艾滋病毒感染具有类似效果的几种精神药物之一。精神药物可能会加速艾滋病毒感染者的抗病毒药物耐药性的发展，这是一个重要的公共卫生问题。

DESCRIPTION (provided by applicant): In the last decade, methamphetamine (METH) use has been correlated with a rising incidence of HIV-1 infection. Furthermore, the reported cases of rapidly progressive HIV-1 infection in METH users underscore the importance of determining the influence of METH on infection of the CNS by HIV-1. The purpose of this grant is to test the hypothesis that METH (and possibly other psychostimulatory drugs) enhances the development of antiviral drug resistant mutants of HIV through a process that accelerates the frequency of infection events, and the concomitant incidence of reverse transcription errors. The emergence of drug resistant genotypes of HIV is prone to take place in the central nervous system (CNS) where drug concentrations may be suboptimal for the effective control of HIV replication. At the same time, the CNS is thought to be under less surveillance by the immune system because of restrictions in T-cell trafficking created by the blood-brain-barrier (BBB). Thus, the CNS serves as a privileged site for HIV infection, replication, adaptation and selection. Furthermore, the CNS may be a reservoir for drug-resistant variants that can enter the peripheral compartment. The proposal has two Specific Aims. Specific Aim 1 is to test the hypothesis that METH accelerates the development of drug-resistant lentiviruses in vitro. This Aim has four objectives: #1: Compare the rate of development of AZT resistant FIV for astrocytes and microglia cultured in the presence or absence of METH. Objective #2: Compare the rate of development in vitro of AZT-resistant HIV for astrocytes and microglia cultured in the presence or absence of METH. Objective #3: Measure the effect of METH on inflammatory cytokine production by uninfected and lentivirus-infected lymphocytes, astrocytes and microglia and correlate with virus expression in these cell types. Objective #4: Determine if METH enhances HIV infection of human target cells, promotes cell proliferation and modulates CXCR4 and/or CCR5 expression. Specific Aim #2 is designed to test the hypothesis in vivo via a proof-of-principle study in the FIV/cat system to evaluate the effect of METH on the development of AZT resistance in vivo. The prospect that psychostimulatory drugs, which include a broad range of legal and illegal substances, can alter the pathogenesis of HIV and influence the emergence of drug-resistant variants is novel. The idea that HIV sequestered in the CNS may have more latitude to become drug-resistant has been proposed previously, but the possible role of METH in this process is a new aspect of that concept. PROJECT NARRATIVE: The CNS is a reservoir for lentivirus infection that is distinct from the peripheral compartment and that allows for the independent evolution of virus variants, especially antiviral drug resistant mutants. Based on our observations, we hypothesize that METH may potentiate the development of drug resistant viruses in the CNS, which then can enter the peripheral compartment. The recent report of a rapidly progressive, highly drug resistant HIV-1 infection in a METH user in New York underscores the importance of elucidating the interactions between METH and HIV-1 and the development of drug resistance in the CNS. In addition, METH is one of several psychotropic drugs that may have similar effects on HIV infection of the CNS. The possibility that psychotropic drugs may accelerate the development of antiviral drug resistance in HIV-infected people is an important public health concern.